We are requesting funds to purchase a PixCell II Laser Capture Microdissection (LCM) System for use by 11 independent investigators, their respective post-doctoral fellows, graduate students, and research technicians. LCM is a new method used to select and procure individual cells or cell clusters from tissue sections. Once captured, the DNA, RNA or protein can be easily extracted from the isolated cells and analyzed by conventional PCR, RT-PCR, or PAGE, including protein zymography. In LCM, a thermoplastic polymer coating [ethylene vinyl acetate (EVA)} is attached to a rigid support that is then placed in contact with a tissue section. The EVA polymer over microscopically selected cell clusters is precisely activated by a near-infrared laser pulse and then bonds to the targeted area. Removal of the EVA and its support from the tissue section procures the selected cell(s) for molecular analysis. This initial NIH LCM approach using a flat transfer EVA film has been recently commercialized Arcturus, Inc., in 1997, and proven to be an effective and routine microdissection technique for subsequent macromolecular analysis. Reliable and precise capture of individual cells from tissue sections, however, was difficult to perform with the initial version of the LCM instrument. Thus, to overcome this limitation, a next generation LCM apparatus was subsequently designed by Arcturus, Inc., the PixCell II, capable of capturing single cells. It is this instrument we wish to purchase. The group of investigators making up this application are from different departments: Cell Biology, Medicine, and Pediatrics. In addition, an investigative group from Johns Hopkins University has joined this application to take advantage of the expertise of the P.I., Dr. Suarez-Quian. During the past year, Dr. Suarez-Quian now wishes to bring this novel technology to Georgetown. The group of investigators assembled have as a central research theme the isolation and molecular fingerprinting of specific cell types from complex tissue. It is precisely the reason for which the PixCell LCM system was developed.